Use of a composition comprising secretions of adipose tissue origin for the manufacture of a medicament for the topical treatment or prevention of acne in an individual
A technique for adipose tissue, topical treatment, applied in the field of use of a composition comprising secretions derived from adipose tissue in the preparation of a medicament for topical treatment or prevention of acne in an individual
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Embodiment 1
[0141] Example 1 Preparation of a mixture of adipocytes and adipose-derived non-adipocytes.
[0142] A 10 g sample of adipose tissue was collected from the inguinal channel of an adult dog by excision. The adipose tissue was rinsed with physiological saline and then completely minced with scissors and mixed with 20 ml of Dulbecco's modified Eagle's medium (DMEM, Sigma). Collagenase (Sigma) was added to a final concentration of 0.05% w / v and samples were incubated at 37°C for 90 minutes. Gently invert samples by hand every 15 minutes during incubation.
[0143] After collagenase treatment, the sample was filtered through a stainless steel mesh (300 μm pore size) under sterile conditions, transferred to a 50 ml centrifuge tube, and centrifuged at 500 g for 15 minutes.
[0144] Four distinct layers are visible within the centrifuged sample: a small (2 mm thick) free lipid layer on the surface, an underlying white 10 mm thick layer of adipocytes, then a large, clear liquid layer...
Embodiment 2
[0147] Example 2 Preparation of cell-free extracts from cell suspensions derived from adipose tissue.
[0148] 10 g of canine adipose tissue samples were completely cut up with scissors, and then mixed with 5 ml of DMEM. Autologous canine serum was filter sterilized and an amount of 1 ml was added to the minced tissue mixture.
[0149] The tissue mixture was incubated overnight at 37°C without agitation. The samples were then centrifuged at 1500 g for 15 minutes, and the fluid between the floating adipocytes and tissue layer and the adipose tissue-derived non-adipocyte pellet was carefully collected. Cell-free extracts contain secretions from adipose tissue.
Embodiment 3
[0150] Example 3 Preparation of another cell-free extract from a cell suspension derived from adipose tissue.
[0151] A 10 g adipose tissue sample was collected from the tail base of the horse by excision. The adipose tissue was rinsed with physiological saline, then roughly cut into small pieces with a diameter of about 5 mm with scissors and mixed with 20 ml of Dulbecco's modified Eagle medium (DMEM, Sigma). Collagenase (Sigma) was added to a final concentration of 0.05% w / v and the samples were incubated at 37°C for 30 minutes, at the end of 30 minutes the adipose tissue was partially digested and composed of partially digested fat particles, free stromal vascular cells (SVC) and a mixture of free adipocytes.
[0152] Samples were then washed by centrifugation at 500g for 15 minutes to remove collagenase. Four distinct layers are visible within the centrifuged sample: a small (2 mm thick) free lipid layer on the surface, a white 10 mm thick layer of adipocytes underneath...
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